肾虚是AD病机之本。补肾名方六味地黄丸在临床应用广泛，其中熟地、山茱萸、山药被谓之方中“三补”。“三补”各单味药有延缓衰老和神经保护作用，但配伍作用机制尚不清楚。秀丽线虫寿命短，基因序列明确，有相关AD模型、IIS通路突变体供研究使用。脑内的IIS通路异常可出现在AD早期，并通过影响Aβ毒性、Tau蛋白过度磷酸化在AD的病理形成、发展中起重要作用。我们前期研究发现，六味地黄丸方可能通过调控IIS通路延长线虫寿命。为进一步探讨其药效机制，本项目拟从AD模型线虫表现型变化入手，应用免疫荧光染色、实时荧光定量PCR、免疫印迹等技术逐步锁定“三补”作用靶点，通过IIS通路相关突变体线虫实验初步证实；并进一步对AD模型线虫进行靶基因RNA干扰，探析“三补”调控IIS通路与AD病理的相关性，阐释其神经保护的作用机制，为相关补肾抗衰中药复方在临床上的应用提供科学依据。

Kidney deficiency is essential in AD pathogenesis. The famous kidney-reinforcing formula, Liuwei Dihuang Pill, has a wide clinical application. Among the herbs that compose the time honored formula, Prepared rehmannia root, Cornus officinalis and Rhizoma dioscoreae are named " three supplement methods" of the formula. They exert anti-aging and neuroprotective effects respectively, but the mechanisms of co-use of them are still undetermined. C. elegans has short lifespan with clear genetic background. Relevant AD model nematode and Insulin/IGF-1 signaling (IIS) pathway mutant options are available for research. The IIS pathway may appear abnormal in early stage of AD. It plays an important role in the formation and development of AD via interacting with β-amyloid toxicity and Tau hyperphosphorylation. Our previous studies observed the anti-aging effect on C.elegans with Liuwei Dihuang Pill. For furthering exploring the underlying mechanisms, this study will observe the changes of phenotype of AD model nematode to target the site in which the "three supplement methods" effect via immunofluorescent staining, real-time fluorescent quantitative PCR, Western blotting, and make initial verification by using IIS pathway relevant transgenic C.elegans. For elucidating the neuroprotective mechanisms of the " three supplement methods ", this study will further interfering RNA of target gene of AD model nematode for investigating the correlation of " three supplement methods " regulating IIS pathway and AD pathology. The outcomes of this study aim at providing scientific basis for clinical use of relevant compound functioning in tonifying kidney with anti-aging effect.